RNA library amplification

• Assemble each library amplifcation reaction as follows:

Component	Volume
Purified, adapter-ligated DNA	10 ul
Library Amplification Master Mix	15 ul
–Total Volume–	–25 ul–

• Mix well by pipetting up and down several times
• Amplify the library using the following thermal cycling protocol:

Step	Temp	Duration	Cycles
Initial denaturation	98 °C	45 sec	1
Denaturation	98 °C	15 sec	12
Annealing-	60 °C	30 sec	12
Extension	72 °C	30 sec	12
Final Extension	72 °C	5 min	1
Hold	10 °C	∞	1

• Place the plate/tube on ice and proceed to –Library Amplification Cleanup–